van Santen Victor J B, Jin Jianfeng, Hogervorst Jolanda M A, Bakker Astrid D
Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam Movement Sciences, Amsterdam, the Netherlands.
Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam Movement Sciences, Amsterdam, the Netherlands.
Biochem Biophys Res Commun. 2023 Feb 26;646:70-77. doi: 10.1016/j.bbrc.2023.01.066. Epub 2023 Jan 21.
Once prostate cancer (PC) metastasizes towards bone the 5-year survival rates drop with 70%, but it is largely unknown why. Bone is continuously mechanically loaded, which likely modulates the paracrine signaling from osteocytes towards PC cells to affect tumor behavior. We hypothesize that shear loaded osteocytes affect PC cell proliferation, invasion and epithelial and mesenchymal-related gene and protein expression. We cultured human DU145 cells, a commonly used cell line for prostate cancer metastases, in the conditioned medium (CM) from shear loaded or unloaded human osteocyte-like-cells (OCYLCs) for 1 and 3 days and assessed their number by staining nuclei with DAPI, their invasion by performing an invasion assay, and epithelial-to-mesenchymal (EMT)-related gene and protein expression by qPCR and immunocytochemistry. CM of shear loaded OCYLCs did not affect DU145 cell number compared to CM of static cultured OCYLCs, but decreased their invasion 1.34-fold. CM of shear loaded OCYLCs enhanced expression of epithelial genes: SYND1 and CDH1 after day 1, while it also enhanced CDH1 after day 3. CM of shear loaded osteocytes enhanced mesenchymal genes: VMN, Snail and MIP2 after day 1, while it decreased expression of mesenchymal CYR61 after day 3. We conclude that CM of shear loaded OCYLCs does not affect DU145 cell proliferation, but decreases their invasion, and differentially affects their EMT-related gene expression. Identifying paracrine signals from shear loaded osteocytes that decrease PC cell invasion may provide novel leads in developing treatments for bone metastases from PC.
一旦前列腺癌（PC）发生骨转移，其5年生存率会下降70%，但其中的原因很大程度上尚不明确。骨骼持续承受机械负荷，这可能会调节从骨细胞到PC细胞的旁分泌信号，从而影响肿瘤行为。我们推测，承受剪切力的骨细胞会影响PC细胞的增殖、侵袭以及上皮和间充质相关基因及蛋白质的表达。我们将人DU145细胞（一种常用于前列腺癌转移研究的细胞系）在来自承受剪切力或未承受剪切力的人骨细胞样细胞（OCYLCs）的条件培养基（CM）中培养1天和3天，通过用DAPI对细胞核进行染色来评估细胞数量，通过侵袭实验评估细胞侵袭能力，并通过qPCR和免疫细胞化学检测上皮-间充质转化（EMT）相关基因及蛋白质的表达。与静态培养的OCYLCs的CM相比，承受剪切力的OCYLCs的CM对DU145细胞数量没有影响，但使细胞侵袭能力降低了1.34倍。承受剪切力的OCYLCs的CM在第1天后增强了上皮基因SYND1和CDH1的表达，在第3天后也增强了CDH1的表达。承受剪切力的骨细胞的CM在第1天后增强了间充质基因VMN、Snail和MIP2的表达，而在第3天后降低了间充质基因CYR61的表达。我们得出结论，承受剪切力的OCYLCs的CM不影响DU145细胞的增殖，但会降低其侵袭能力，并对其EMT相关基因表达产生不同影响。识别来自承受剪切力的骨细胞的降低PC细胞侵袭能力的旁分泌信号，可能为开发前列腺癌骨转移的治疗方法提供新的线索。
